Anti-CD63 antibody [EPR22458-280]

• Flow Cyt

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Flow Cytometry - Anti-CD63 antibody [EPR22458-280] (AB252919)

Flow cytometric analysis of SK-MEL-28 (human malignant melanoma cell line) cells labeling CD63 with ab252919 at 1/60 (red) compared with a Rabbit monoclonal IgG Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.

Left panel : No fixation/permeabilization; Gated on viable cells for surface CD63 staining.

Right panel : 4% paraformadehyde fixation / 90% methanol permeabilization.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CD63 antibody [EPR22458-280] (AB252919)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SK-MEL-28 (human malignant melanoma cell line) cells labeling CD63 with ab252919 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing lysosome/endosomes co-staining in SK-MEL-28 cell line. The nuclear counterstain is DAPI (blue). Lysosomes/endosomes are detected with Anti-LAMP1 antibody [H4A3] (ab25630) at 1/200 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed (ab150120) (red).

Secondary antibody only control : Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CD63 antibody [EPR22458-280] (AB252919)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SK-MEL-28 (human malignant melanoma cell line) cells labeling CD63 with ab252919 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and membranous staining in SK-MEL-28 cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-CD63 antibody [EPR22458-280] (AB252919)

Flow cytometric analysis of human peripheral blood mononuclear cells labeling CD63 with ab252919 at 1/60 (right) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (left). Goat Anti-Rabbit IgG Fc (Alexa Fluor^® 488) preadsorbed (ab150097), at 1/5000 dilution was used as the secondary antibody.

Cells were stained with rabbit IgG (Left) or ab252919 (Right). Then stained with anti-CD14 conjugated to Alexa Fluor^® 647.

Gated on viable cells.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD63 antibody [EPR22458-280] (AB252919)

Immunohistochemical analysis of paraffin-embedded human breast tissue labeling CD63 with ab252919 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Counter stained with hematoxylin.
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Negative control : No staining on normal epithelial cells and stromal cells in human breast (PMID : 22957045).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD63 antibody [EPR22458-280] (AB252919)

Immunohistochemical analysis of paraffin-embedded human clear cell renal cell carcinoma tissue labeling CD63 with ab252919 at 1/4000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101), Ready to use. Counter stained with hematoxylin. Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins. Positive staining on human clear cell renal cell carcinoma. The section was incubated with ab252919 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD63 antibody [EPR22458-280] (AB252919)

Immunohistochemical analysis of paraffin-embedded human melanoma tissue labeling CD63 with ab252919 at 1/2000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human melanoma (PMID : 22425483) is observed. Counter stained with hematoxylin.
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 minutes.

The section was incubated with ab252919 for 30 minutes at RT.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.